Proteomic Analysis of GLUT4 Storage Vesicles Reveals Tumor Suppressor Candidate 5 (TUSC5) as a Novel Regulator of Insulin Action in Adipocytes

• Published in: The Journal of biological chemistry Vol. 290; no. 39; pp. 23528 - 23542
• Main Authors: Fazakerley, Daniel J., Naghiloo, Sheyda, Chaudhuri, Rima, Koumanov, Françoise, Burchfield, James G., Thomas, Kristen C., Krycer, James R., Prior, Matthew J., Parker, Ben L., Murrow, Beverley A., Stöckli, Jacqueline, Meoli, Christopher C., Holman, Geoffrey D., James, David E.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 25.09.2015; American Society for Biochemistry and Molecular Biology
• Subjects: 3T3-L1 Cells; adipocyte; Adipocytes - physiology; Animals; Cell Biology; glucose transporter type 4 (GLUT4); Glucose Transporter Type 4 - metabolism; insulin; Insulin - physiology; insulin resistance; Male; membrane trafficking; Mice; Proteomics; Rats; Rats, Wistar; tumor suppressor candidate 5 (TUSC5); Tumor Suppressor Proteins - genetics; Tumor Suppressor Proteins - physiology; glucose transporter type 4 (GLUT4); tumor suppressor candidate 5 (TUSC5); insulin; insulin resistance; membrane trafficking; adipocyte
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M115.657361

Insulin signaling augments glucose transport by regulating glucose transporter 4 (GLUT4) trafficking from specialized intracellular compartments, termed GLUT4 storage vesicles (GSVs), to the plasma membrane. Proteomic analysis of GSVs by mass spectrometry revealed enrichment of 59 proteins in these vesicles. We measured reduced abundance of 23 of these proteins following insulin stimulation and assigned these as high confidence GSV proteins. These included established GSV proteins such as GLUT4 and insulin-responsive aminopeptidase, as well as six proteins not previously reported to be localized to GSVs. Tumor suppressor candidate 5 (TUSC5) was shown to be a novel GSV protein that underwent a 3.7-fold increase in abundance at the plasma membrane in response to insulin. siRNA-mediated knockdown of TUSC5 decreased insulin-stimulated glucose uptake, although overexpression of TUSC5 had the opposite effect, implicating TUSC5 as a positive regulator of insulin-stimulated glucose transport in adipocytes. Incubation of adipocytes with TNFα caused insulin resistance and a concomitant reduction in TUSC5. Consistent with previous studies, peroxisome proliferator-activated receptor (PPAR) γ agonism reversed TNFα-induced insulin resistance. TUSC5 expression was necessary but insufficient for PPARγ-mediated reversal of insulin resistance. These findings functionally link TUSC5 to GLUT4 trafficking, insulin action, insulin resistance, and PPARγ action in the adipocyte. Further studies are required to establish the exact role of TUSC5 in adipocytes. Background: We searched for novel regulators of insulin-stimulated glucose transport in adipocytes. Results: Tumor suppressor candidate 5 (TUSC5) colocalized with GLUT4, and manipulation of TUSC5 expression levels affected insulin-regulated glucose transport. Conclusion: TUSC5 is a novel regulator of insulin-stimulated glucose transport. Significance: TUSC5 contributes to insulin-sensitizing effects of PPARγ agonists in adipocytes.